A scalp care composition with bacteriostatic, anti-dandruff and anti-itching effects, its preparation method and application

Abstract

The present invention belongs to the field of biotechnology. More specifically, it relates to a scalp care composition having the effects of inhibiting bacteria, removing dandruff and relieving itching, and its preparation method and application. In particular, the composition contains the extract of Alpinia katsumadai and piroctone olamine. The extract of Alpinia katsumadai in the present invention has good antibacterial activity against Malassezia globosa. The combined drug sensitivity test shows that the combination of the extract of Alpinia katsumadai and piroctone olamine has a synergistic effect in inhibiting Malassezia globosa.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology. More specifically, it relates to a scalp care composition having antibacterial, anti-dandruff and antipruritic effects, its preparation method and application. In particular, the composition contains the extract of Alpinia katsumadai and piroctone olamine. Background Art

[0002] Malassezia mainly grows in the stratum corneum of human skin and is related to the triglycerides and free fatty acids of the sebaceous glands present on the skin surface. It can cause various skin diseases such as pityriasis versicolor, Malassezia folliculitis, seborrheic dermatitis and dandruff. At present, the drugs for inhibiting Malassezia mainly include chemical antifungal agents such as ketoconazole, clotrimazole, zinc pyrithione, etc. However, most of them are chemically synthesized compounds, and the long-term use of such substances may cause the patients to have dry scalp, excessive dandruff, and be prone to drug resistance.

[0003] Alpinia katsumadai is the dried nearly mature seeds of the perennial herb Alpinia katsumadai of the genus Alpinia in the family Zingiberaceae, and has the effects of drying dampness and promoting qi movement, warming the middle-jiao and stopping vomiting. Research shows that the volatile oil in Alpinia katsumadai has the effects of protecting gastric mucosa, anti-gastric ulcer, anti-inflammatory, antibacterial, etc. [1-3] . Wei Jieshu et al. [4] found that the volatile oil of the seed mass of Alpinia katsumadai has a certain inhibitory effect on Escherichia coli, Staphylococcus aureus and Bacillus subtilis. Peng Fu et al. [5] confirmed that the volatile oil of Alpinia katsumadai can inhibit the pathogens of cow mastitis. In addition, Chu Zuochen [6] found that the volatile oil microemulsion of Alpinia katsumadai has a good inhibitory effect on banana anthracnose and has a certain fresh-keeping effect on bananas. However, there is no report on the inhibitory effect of the extract of Alpinia katsumadai on Malassezia.

[0004] [1] Xie Peng, Qin Huazhen, Tan Ximei, etc. Research progress on chemical constituents and pharmacological effects of Alpinia katsumadai [J]. Journal of Liaoning University of Traditional Chinese Medicine, 2017, 19(3): 60-63.

[0005] [2] Wu Zhen, Chen Yongshun, Du Shiming, etc. Effects of volatile oil of Alpinia katsumadai on acetic acid-induced gastric ulcer in rats [J]. Chinese Journal of Hospital Pharmacy, 2010, 30(7): 560-563.

[0006] [3]BARBARAG,OLAFK,FRANZB.CompoundsofAlpiniakatsumadaiaspotentialeffluxinhibitorsinMycobacteriumsmegmatis[J].Bioorganic&MedicinalChemistry,2012,20(8):2701-2706.

[0007] [4]Weijieshu,Hujinshan,HongQunhua,etal.StudyontheAntibacterialActivityofVolatileOilsfromAlpiniavillosaandAlpiniaKatsumadaiHayata[J].BiologicalChemicalEngineering,2023,9(3):45-50.

[0008] [5]PengFu,DaiMin,WanFeng.StudyontheAntibacterialActivityofEffectivePartsofAlpiniaKatsumadaiHayataagainstPathogenicBacteriainDairyCowswithMastitis[J].ChineseVeterinaryScience,2012,42(10):1073-1080.

[0009] [6]ChuZuochen.PreparationofMicroemulsionofAlpiniaKatsumadaiHayataEssentialOilandItsEffectonBananaFresh-keeping[D].Wuhan:HuazhongAgriculturalUniversity,2022. SUMMARYOFTHEINVENTION

[0010] (1)ItisdiscoveredthroughresearchthatthespecificsolventextractionpartofthealcoholicextractofAlpiniaKatsumadaiHayatahasasignificantinhibitoryeffectonMalasseziaglobosa,andtheeffectismostobviouscomparedwithotherextractionparts.Furtherstudyonitsinteractionwithtraditionalanti-dandruffagentsrevealsthattheextractofAlpiniaKatsumadaiHayataonlyshowssynergisticeffect(FICI≤0.5)withpiroctoneethanolaminesalt,additiveeffectwhencombinedwithketoconazoleandclimbazole,andirrelevanteffectwhencombinedwithZPTandHD-100.

[0011] ThepurposeofthisinventionistoprovidetheuseoftheextractofAlpiniaKatsumadaiHayatainthepreparationofaninhibitoragainstMalassezia.

[0012] AnotherpurposeofthisinventionistoprovidetheuseoftheextractofAlpiniaKatsumadaiHayatainthepreparationofamedicinefortreatingskindiseasescausedbyMalasseziainfection.

[0013] Inoneembodimentofthisinvention,theskindiseasesinclude(tineaversicolor)andseborrheicdermatitis.

[0014] AnotherpurposeofthisinventionistoprovidetheuseoftheextractofAlpiniaKatsumadaiHayatainthepreparationofaninhibitory,anti-dandruffandanti-itchingscalpcareproduct.

[0015] Inoneembodimentofthisinvention,theMalasseziaisMalasseziaglobosa.

[0016] Inoneembodimentofthisinvention,theextractofAlpiniaKatsumadaiHayataisthen-hexaneextractionpartofthealcoholicextractofAlpiniaKatsumadaiHayata.

[0017] Another object of the present invention is to provide an antibacterial, anti-dandruff and anti-itching scalp care composition, which contains Alpinia katsumadai extract at a concentration of about 25 - 150 μg / mL and piroctone olamine at a mass of about 0.1 - 1.0%.

[0018] In one embodiment of the present invention, the composition further comprises: a pharmaceutically and / or cosmetically acceptable carrier.

[0019] In one embodiment of the present invention, the pharmaceutically and / or cosmetically acceptable carrier comprises: a combination of two or more of surfactants, humectants, chelating agents, emollients, pH regulators and fragrances.

[0020] In one embodiment of the present invention, the composition is made in the form of: shampoo, hair conditioner, hair essence, hair paste or hair gel.

[0021] The present invention has the following beneficial effects:

[0022] (1) The results of the antibacterial zone test and MIC value test show that the diameter of the antibacterial zone of the Alpinia katsumadai extract of the present invention against Malassezia globosa is > 20 mm, and the antibacterial effect is the most obvious, being extremely sensitive.

[0023] (2) The results of the combined drug sensitivity test show that the combination of the Alpinia katsumadai extract and traditional anti-dandruff agents does not all show a synergistic effect. Only the combination with OCT shows a synergistic effect, with FICI ≤ 0.5. The combinations with ketoconazole and clotrimazole show an additive effect, and the combinations with ZPT and HD-100 show an irrelevant effect. Detailed Embodiments

[0024] The following specific examples are used to further illustrate the present invention, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the technical field.

[0025] Unless otherwise specified, the reagents and materials used in the following examples are all commercially available.

[0026] Among them, the solid medium for Malassezia: 1% peptone, 2% agar, 1% glucose, 0.2% yeast extract, 0.05% glycerol monostearate, 0.5% Tween-60, 1% glycerol, 0.8% bile salts and 2% olive oil, made up to 1000 mL, pH 7.0 - 7.2; the liquid medium does not contain agar.

[0027] Experimental Example 1: Comparison of the antibacterial activities of different extraction parts of Alpinia katsumadai

[0028] 1.1 Preparation of different extraction parts of Alpinia katsumadai: Take the dry seeds of Alpinia katsumadai, crush them into coarse powder and sieve. Add ethanol solution with a volume fraction of 65% according to the material-liquid ratio of 1:10, soak for 30 min, then heat under reflux at 70 °C for 2 times, each time for 1.5 h. Collect the extraction solution, filter to remove the medicinal residues. Concentrate the filtered liquid, refrigerate to precipitate ethanol, recover the precipitated ethanol, dry the supernatant to obtain dry extract. Add 5 times the weight of n-hexane, petroleum ether, chloroform and ethyl acetate to the above dry extract respectively, extract at 45 °C for 2.0 h, concentrate under reduced pressure to constant weight and dry to obtain the extraction parts of n-hexane, petroleum ether, chloroform and ethyl acetate respectively for standby.

[0029] 1.2 Test method

[0030] 1.2.1 Activation of strains and preparation of bacterial suspensions: The tested strains (Malassezia globosa CBS9757, purchased from the Medical Fungal Preservation Center of the Institute of Dermatology, Chinese Academy of Medical Sciences) were subcultured on the solid medium of Malassezia for 2 times. Pick single colonies and inoculate them into the liquid medium of Malassezia, and incubate at a constant temperature of 32 °C in a shaker at 220 r / min for 48 h. Use the method of double dilution of bacterial liquid to prepare a bacterial liquid with a final concentration of 1×10 6 CFU / mL as the experimental bacterial suspension.

[0031] 1.2.2 Preparation of medicinal liquids: Dissolve the extraction parts of n-hexane, petroleum ether, chloroform and ethyl acetate with dimethyl sulfoxide respectively to prepare medicinal liquids with a mass concentration of 100 mg / mL, and store them at 4 °C for standby; the concentration of the positive drug octopirox ethanolamine salt (OCT) is 50 mg / mL.

[0032] 1.2.3 Antibacterial circle test: According to the procedure reported by the Clinical and Laboratory Standards Institute (CLSI), use the Kirby-Bauer disk diffusion method to detect the antibacterial activity of each medicinal liquid. Use a pipette to suck 200 μL of the bacterial suspension onto the culture medium plate, spread it evenly and set aside. Drop 20 μL of the medicinal liquid on a sterile circular filter paper with a specification of (6.0 mm × 1.0 mm). Gently stick the filter paper on the surface of the agar plate with tweezers. After incubating the petri dish at a constant temperature of 37 °C for 48 h, observe and measure the diameter of the antibacterial circle, and take its average value as the measurement result. The edge of the antibacterial ring is limited to the point where no obvious growth of bacteria can be seen with the naked eye. The result determination is based on the "Performance Standards for Antimicrobial Susceptibility Testing" promulgated by the National Committee for Clinical Laboratory Standards (NCCLS) of the United States: a diameter > 20 mm is extremely sensitive, 15 - 20 mm is highly sensitive, 10 - 15 mm is moderately sensitive, and < 10 mm is lowly sensitive.

[0033] 1.3 Determination of MIC value: The reference standard is the M27 standard - broth dilution antifungal susceptibility test reference method for yeasts formulated by the Clinical and Laboratory Standards Institute (CLSI). The drug solution was diluted by a factor of two in 10 gradients using distilled water. A 96-well plate was taken, and 96 μL of liquid medium was added to columns 1-11. 4 μL of a single drug solution in 10 gradients was added to column 1. The drug solution concentrations decreased from high to low. 100 μL of bacterial solution was added to each well in columns 1-11. Three parallels were set for each drug. Column 11 was used as the growth control, and an OCT positive control group and a negative control group were also set. After incubating the inoculated 96-well plate at 30 °C for 48 h, the results were observed. The positive wells showed a color change from yellow to red, and the lowest drug concentration at which the fungus was completely inhibited (no color change) in that well was the MIC.

[0034] 2 Results and Discussion

[0035] 2.1 Results of inhibition zone test

[0036] The results of the inhibition zone are shown in Table 1 below. The inhibition zone diameter of the n-hexane fraction of Alpinia katsumadai against Malassezia globosa at 50 mg / mL was > 20 mm, and the antibacterial effect was the most obvious, being extremely sensitive; the inhibition zone diameter of the chloroform fraction of Alpinia katsumadai against Malassezia at the same concentration was < 10 mm; the petroleum ether extraction fraction had a moderately sensitive antibacterial effect on Malassezia, while the ethyl acetate fraction had no antibacterial activity.

[0037] Table 1 Comparison of inhibition zone diameters of different extraction fractions against Malassezia globosa

[0038] Liquid medicine Diameter of inhibition zone (mm) Hexane fraction of Alpinia katsumadai 32.62±2.13 Petroleum ether fraction of Alpinia katsumadai <![CDATA[13.52±1.03 ** > Chloroform fraction of Alpinia katsumadai <![CDATA[8.29±0.75 *** > Ethyl acetate fraction of Alpinia katsumadai _***

[0039] Compared with the n-hexane fraction of Alpinia katsumadai, ** P < 0.01, *** P < 0.001.

[0040] 2.2 Results of MIC test

[0041] Table 2 Results of MIC test of the n-hexane fraction of Alpinia katsumadai against Malassezia globosa

[0042] Liquid medicine MIC (μg / mL) Hexane fraction of Alpinia katsumadai 125 OCT 62.5

[0043] Experimental Example 2: Combined drug susceptibility test

[0044] The checkerboard dilution method was used for the combined drug sensitivity test, and the fractional inhibitory concentration index (FICI) was measured to evaluate the combined drug effect of the n-hexane fraction of Alpinia katsumadai (hereinafter referred to as the Alpinia katsumadai extract) and other drugs. The maximum stock solution concentration of the Alpinia katsumadai extract was 100 mg / mL, the maximum stock solution concentrations of piroctone olamine (OCT) and hydroxyethyl disulfonate (HD-100) were 50 mg / mL, and the maximum stock solution concentrations of ketoconazole, zinc pyrithione (ZPT), and clotrimazole were 25 mg / mL.

[0045] The drug solution was diluted by two-fold dilution method with distilled water for 10 gradients. 96 μL of liquid medium was added to columns 1-10 of the 96-well plate. 2 μL of the two-fold concentration drug solution of each of the two drugs was added horizontally and vertically. Horizontally, there were 10 rows of the 10 concentration gradients of the Alpinia katsumadai extract from high to low, and vertically, there were 10 columns of the 10 concentration gradients of the traditional anti-dandruff agent from high to low, so that different combined drug concentrations were formed in 80 wells of the two drugs. Then, 100 μL of the bacterial solution was added to each well of columns 1-11. Well 11 was used as the positive control, and well 12 was used as the negative control. The 96-well plate was placed at 30 °C for 48 h to observe the results. The positive wells showed a color change from yellow to red. The fractional inhibitory concentration index (FICI) was used to evaluate the combined drug effect of hinokitiol and other drugs. The calculation is shown in the following formula.

[0046]

[0047] Where A and B are the Alpinia katsumadai extract and the traditional anti-dandruff agent respectively, and MICA and MICB are the MIC values of A and B when used alone. MICA 联用 is the MIC value of A when A and B are used in combination, and MICB 联用 is the MIC value of B when A and B are used in combination. When FICI ≤ 0.5, it is a synergistic effect; when 0.5 < FICI ≤ 1, it is an additive effect; when 1 < FICI ≤ 2, it is an irrelevant effect; when FICI > 2, it is an antagonistic effect.

[0048] Table 3 MIC values of different drugs against Malassezia globosa

[0049] Liquid medicine MIC (μg / mL) Extract of Alpinia katsumadai 125 OCT 62.5 Ketoconazole 3.91 ZPT 15.63 Climbazole 7.81 Imidazolidinyl urea 31.25

[0050] Table 4 Test results of the combined drug sensitivity test

[0051]

[0052] As can be seen from the above table, the combined use of the Alpinia katsumadai extract and the traditional anti-dandruff agent does not all show a synergistic effect. It only shows a synergistic effect in the combination with OCT, with FICI ≤ 0.5. The combinations with ketoconazole and clotrimazole both show an additive effect, and the combinations with ZPT and HD-100 show an irrelevant effect.

[0053] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. Use of an extract of cardamom vine in the preparation of an inhibitor of Malassezia sphaerocephala, characterized in that: The cardamom extract is the n-hexane extraction portion of the cardamom ethanol extract.

2. An antibacterial, anti-dandruff and antipruritic scalp care composition, characterized in that: The invention comprises the cardamom extract according to claim 1 with a concentration of 25-150 μg / mL and 0.1-1.0% by weight of piroctone olamine.

3. The scalp care composition according to claim 2, characterized in that The composition further comprises: a pharmaceutically and / or cosmetically acceptable carrier.

4. The scalp care composition according to claim 3, characterized in that The pharmaceutically and / or cosmetically acceptable carrier comprises a combination of two or more of a surfactant, a moisturizer, a chelating agent, an emollient, a pH adjuster and a fragrance.

5. The scalp care composition according to claim 4, characterized in that The composition is prepared in the form of shampoo, conditioner, hair essence, hair cream or hair gel.

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